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The primary objective was to evaluate the efficacy and safety of Er:YAG laser treatment for meibomian gland dysfunction (MGD) in a prospective study. A total of 128 eyes from 64 patients with MGD were enrolled to receive either three Er:YAG laser treatments with meibomian gland expression (MGX) or MGX-alone treatment sessions at 3-week intervals. The Standard Patient Evaluation of Eye Dryness (SPEED) validated questionnaire; fluorescein breakup time of the tear film (FBUT); corneal fluorescein staining (CFS); lid margin abnormalities; meibomian gland morphology (meiboscore); lower tear meniscus height (TMH); and assessment of 15 meibomian glands in the lower eyelids, including total meibomian gland secretion quality (TMGS), the number of glands secreting any liquid (GSAL), and the number of glands yielding optimal clear liquid secretion (GYCL), were assessed at day (D)0, D21, D42, and D63 for the Er:YAG-MGX group and D0 and D63 for the MGX group. At D63, significant decreases in SPEED scores and lid margin abnormalities as well as significant increases in FBUT, TMGS, and GSAL were observed in both groups (all p

The quorum-sensing (QS) signaling-dependent extracellular virulence factors of Pseudomonas aeruginosa can cause infections such as P. aeruginosa keratitis. P. aeruginosa communicates by secreting and sensing small chemical molecules called autoinducers in QS system. The key QS signal molecule, N-3-oxododecanoyl-homoserine lactone (3OC12HSL), can affect the behavior of host cells and initiate immune response. In this report we investigated the influence of 3OC12HSL on human corneal epithelial cells (HCECs) and the mechanisms of 3OC12HSL on activated toll-like receptor 2 (TLR2)-dependent interleukin-8 (IL-8) secretion in HCECs. Cells were cultured under different concentrations of 3OC12HSL. Cell viability was assessed using Crystal violet staining and the cell counting kit-8 assay. We demonstrated the administration of 3OC12HSL decreased HCEC viability and survival in a concentration- and time-dependent manner. At high concentrations, 3OC12HSL rapidly promoted a time-dependent increase in the expressions of TLR2 and TLR4. It was found that the nuclear translocation and expression of nuclear factor-κB (NF-κB) were also increased in response to 3OC12HSL treatment. The significantly elevated expressions of TLR2, TLR4, and NF-κB, encouraged us to further test their mechanisms that cause inflammatory response. Among the inflammatory factors examined (IL-6, IL-8, IL-10, and TNF-α), we found that IL-8 was significantly increased after treatment with 3OC12HSL and its expression was inhibited when TLR2 was specifically blocked or silenced. These results indicated that the QS signaling molecule 3OC12HSL could be recognized by the host innate immune system in HCECs. This recognition then triggered an immune inflammatory response involving the activation of TLR2 and an increase in expression of IL-8. This crosstalk between 3OC12HSL and host immunity in HCECs contributes to the development and progression of P. aeruginosa keratitis.

Novel functional AIEgen based on three compact bound aryl skeletons is designed and synthesized. This tri-aryl type luminogen (TA-Catechol) embedded with catechol moiety responds rapidly to series of boronic acids. Real-time visual and quantitative dual-mode detection method is established for the first time with modest precision and low detection limit (8.0 μM). Detailed mechanistic discussion identifies tetra-coordinated boronic species as the key intermediate within sensing procedure. Wide range of organic boronic acids compatible with this strategy is displayed which is promising in high throughput screening technology. Furthermore, solid-state sensing capability of TA-Catechol is also demonstrated.

A new aggregation-induced emission (AIE) luminescence core, 1-hydroquinol-1,2,2-triphenylethene (HQTPE), has been designed and synthesized for the first time. This AIE core is amazingly simple but is fundamentally important to chemistry because of its intrinsic redox and pH activities. The incorporation of hydroquinone (HQ) moiety into a common AIE core tetraphenylethene (TPE) yields HQTPE with unique fluorescent properties over most other AIEgens so far reported. There are tremendous differences of photochemcical properties between HQTPE, 1-benzoquinol-1,2,2-triphenylethene (QTPE, the oxidized counterpart) and its anions. Interestingly, as the solution concentration is increased, AIEgen HQTPE shows stronger fluorescence but QTPE exhibits rapid quenching of fluorescence in a nonlinear fashion, which are in agreement with theoretical studies. The fluorescence of HQTPE is also highly dependent of pH value of media. We have further explored HQTPE as an ultrasensitive redox probe and efficient deoxidizer, which could lead to many potential applications in health care, food security, environmental monitoring, optic and electronic devices.

At present, the clinical detection method of human papillomavirus (HPV) is mainly based on the PCR method. However, this method can only be used to detect HPV DNA and HPV types, and cannot be used to accurately predict cervical cancer. HPV16 E7 is an oncoprotein selectively expressed in cervical cancers. In this study, we prepared an HPV16 E7-histidine (HIS) fusion oncoprotein by using a prokaryotic expression and gained several mouse anti-HPV16 E7-HIS fusion oncoprotein monoclonal antibodies (mAbs) by using hybridoma technology. Two mAbs, 69E2 (IgG2a) and 79A11 (IgM), were identified. Immunocytochemistry, immunofluorescence, immunohistochemistry, and Western blot were used to characterize the specificity of these mAbs. The sequences of the nucleotide bases and predicted amino acids of the 69E2 and 79A11 antibodies showed that they were novel antibodies. Indirect enzyme-linked immunosorbent assay (ELISA) with overlapping peptides, indirect competitive ELISA, and 3D structural modeling showed that mAbs 69E2 and 79A11 specifically bound to the three exposed peptides of the HPV16 E7 (HPV16 E749&ndash, 66, HPV16 E773&ndash, 85, and HPV16 E791&ndash, 97). We used these two antibodies (79A11 as a capture antibody and 69E2 as a detection antibody) to establish a double-antibody sandwich ELISA based on a horseradish peroxidase (HRP)-labeled mAb and tetramethylbenzidine (TMB) detection system for quantitative detection of the HPV16 E7-HIS fusion oncoprotein, however, it was not ideal. Then we established a chemiluminescence immunoassay based on a labeled streptavidin-biotin (LSAB)-ELISA method and luminol detection system&mdash, this was sufficient for quantitative detection of the HPV16 E7-HIS fusion oncogenic protein in ng levels and was suitable for the detection of HPV16-positive cervical carcinoma tissues. Collectively, we obtained two novel mouse anti-HPV16 E7 oncoprotein mAbs and established an LSAB-lumino-dual-antibody sandwich ELISA method for the detection of the HPV16 E7-HIS fusion oncogenic protein, which might be a promising method for the diagnosis of HPV16-type cervical cancers in the early stage.

AIM: To observe the efficacy of different artificial eye drops on corneal epithelium healing in rabbit. METHODS: Thirty-five rabbits with 6 mm diameter central corneal epithelium removed were randomly assigned to six groups: 0.9% normal saline (NS) group, 0.1% hyaluronate (HA) group, 0.3% HA group, Tears Naturale Free® (TNF) group, 0.4% polyethylene glycol (PEG) group, 0.5% carboxymethyl cellulose (CMC) group and blank control group. Treatments were administered topically four times daily. Corneal epithelium healing was evaluated by the percentage reduction in wound area at 24, 36, 48, 60, and 72h after removal of the corneal epithelium. Cornea re-epithelialization was also assessed by histological analysis and electron microscopy. RESULTS: All corneal wounds completely re-epithelialized in less than 72h. The average re-epithelialization time was 47.61±4.25h in the 0.3% HA group and 49.72±1.05h in the 0.9% NS group, followed by 0.1% HA, TNF, 0.4% PEG, 0.5% CMC, and lastly by the control group. Compared to the control group, there were significant differences among 0.3% HA, 0.9% NS, PEG, and TNF (P

The current research of mechanochromic molecules focused on different molecular skeletons and the types of functional groups, where the design ideas were limited. We have synthesized a group of cyano-modified TPE derivatives (TPECN) with different substitution sites, showing typical aggregation induced emission (AIE) effect. We found that meta-substituted mCN revealed the inertness of mechanochromism, whether being ball milled or rapidly evaporated. Isomers with ortho- or para-substitution showed an emission wavelength shift over 40 nm. Experimental data indicated unusual high-strength lattice stability for mCN. By single-crystal analysis and theoretical calculations, we clarified from the molecular skeleton distortion, noncovalent intermolecular interactions, and calculated interaction energy that the meta-substitution led to a stable lattice structure, making mCN insensitive to external stimuli. Thus, we successfully explained the substituted site was an important and independent factor that influenced the mechanochromic property. Meanwhile, we also explored the application of the TPECNs series for erasable printing and coding.

Catalytic oxidative cross-dehydrogenative coupling between unactivated C(sp2)–H and C(sp3)–H bonds is achieved by the cobalt-catalyzed o-alkylation reaction of aromatic carboxamides containing (pyridin-2-yl)isopropyl amine (PIP–NH2) as a N,N-bidentate directing group. Many different C(sp3)–H bonds in alkanes, toluene derivatives and even in the α-position of ethers and thioethers can be used as coupling partners. This method has a broad substrate scope and the tolerance of various functional groups.

The first cobalt-catalyzed direct methylation of a C(sp(2) )-H bond using dicumyl peroxide (DCP) as both the methylating reagent and hydrogen acceptor has been established. The reaction proceeded without the use of any additives, and was proven to be applicable to various amides bearing a 2-pyridinylisopropyl (PIP) directing group, providing an efficient access to o-methyl aryl amides with high functional-group tolerance. Preliminary mechanistic studies suggest a radical process would be involved in the catalytic process.

Objectives To investigate the effects of neutrophil extracellular traps (NETs) on angiogenesis in vitro and in vivo and the regulatory role of mammalian target of rapamycin (mTOR) activity in it. Methods The regulatory role of mTOR in NETs formation was explored. In vitro, human neutrophils were pretreated with rapamycin. NETs formation was measured using immunofluorescence staining of NETs markers, SYTOX Green and PicoGreen after NaOH stimulation. In vivo, mice were treated with rapamycin, and NETs formation in cornea was measured using immunofluorescence staining 7 days after alkali burn. Then, the effects of NETs on angiogenesis were investigated. In vitro, human neutrophils were treated with DNase I or rapamycin. NETs were isolated after NaOH stimulation and the isolated NETs were co-culture with human umbilical vein endothelial cells (HUVECs). HUVECs migration, proliferation, and inflammatory activation were measured. In vivo, mice were injected subconjunctivally with supernatant containing NETs. Corneal neovascularization was visualized by immunofluorescence staining. Results NETs structures can be observed in NaOH-stimulated neutrophils and alkali-burned mouse cornea compared with normal group. Treated with rapamycin enhanced NETs formation in response to NaOH management compared with DMSO control in vitro and in vivo. NETs increased the migration, proliferation and inflammatory activation of HUVECs, and subconjunctival injection of NETs promoted inflammatory and angiogenic response in corneal alkali burn model. Conclusions NETs formation can be triggered by NaOH stimulation. mTOR activity has a negative regulatory effect on NETs formation. NETs promoted angiogenic responses and inflammatory activation of HUVECs and increased corneal neovascularization and inflammatory response.

Glioma is the most common and malignant form of primary brain tumour, and is characterised by high proliferation and extensive invasion and neurological destruction. Demethylzeylasteral (T-96), which is extracted from Tripterygium wilfordii, is considered to have immunosuppressive, anti-inflammatory and anti-angiogenic effects. Here, the anti-tumour effect of T-96 on glioma was evaluated. Our results demonstrated that T-96 significantly inhibited glioma cell growth and induced cell cycle arrest in G1 phase but did not induce apoptosis. Cell invasion and migration were dramatically suppressed after treatment with T-96. Almost all genes related to cell cycle and DNA replication were downregulated after treatment with T-96. Our results showed that miR-30e-5p was noticeably upregulated after T-96 treatment, and MYBL2, which is involved in cell cycle progression and is a target gene of miR-30e-5p, was significantly reduced in synchrony. Overexpression of MYBL2 partially rescued the T-96-induced inhibition of cell growth and proliferation. Moreover, a miR-30e-5p antagomir significantly reduced the upregulation of miR-30e-5p expression induced by T-96, leading to recovery of MYBL2 expression, and partially rescued the T-96-induced inhibition of cell growth and proliferation. More important, T-96 effectively upregulated miR-30e-5p expression and downregulated MYBL2 expression, thus inhibiting LN-229 cell tumour growth in a mouse model. These results indicated that T-96 might inhibit glioma cell growth by regulating the miR-30e-5p/MYBL2 axis. Our study demonstrated that T-96 might act as a promising agent for malignant glioma therapy.

Triptolide is a diterpene triepoxide, extracted from the Chinese herb Tripterygium wilfordii Hook F, which has been shown to have antitumor activity in a number of cancers. Neuroblastoma is an aggressive extracranial pediatric solid tumor, with significant chemotherapeutic resistance. In this study, triptolide was hypothesized to be a potential therapeutic agent for neuroblastoma. The effects of triptolide on neuroblastoma cell growth and tumor development were investigated. Cell growth and proliferation were evaluated using a cell counting kit‑8 assay and a 5-bromo-2-deoxyuridine staining assay. Cell cycle and apoptosis were detected by flow cytometry. Reverse transcription‑quantitative polymerase chain reaction was conducted to detect the expression levels of the apoptosis‑associated proteins, caspase‑3 and caspase‑9. The tumorigenicity of neuroblastoma cells was assessed by a soft agar clonogenic assay and an in vivo tumorigenic assay. The results demonstrated that exposure of BE(2)‑C human neuroblastoma cells to triptolide resulted in a reduction in cell growth and proliferation, and the induction of cell death and apoptosis, together with cell cycle arrest in the S phase. A soft agar assay indicated that triptolide inhibited the colony‑forming ability of BE(2)‑C neuroblastoma cells. The xenograft experiment showed that triptolide significantly reduced tumor growth and development in vivo. The data suggested that this Chinese herb may be a potential novel chemotherapeutic agent for neuroblastoma.

Neutrophils release neutrophil extracellular traps (NETs) in a pathogen-killing process called NETosis. Excessive NETs formation, however, is implicated in disease pathogenesis. Therefore, to understand how NETosis is regulated, we examined the effect of dexamethasone (DXM), an anti-inflammatory drug, on this process and the role of toll-like receptors (TLRs). We stimulated human neutrophils with phorbol 12-myristate 13-acetate (PMA) or Staphylococcus aureus (S. aureus) and quantified NETs formation. We also examined the effect of DXM on the bactericidal effect of NETs and the role of reactive oxygen species (ROS) and nuclear factor (NF)-κB in DXM-regulated NETosis. DXM significantly inhibited S. aureus-induced NETosis and extracellular bacterial killing. ROS production and NF-κB activation were not involved in DXM-regulated NETosis. TLR2 and TLR4, but not TLR5 or TLR6, modified S. aureus-induced NETs formation. Neither DXM nor TLRs were involved in PMA-induced NETosis. Furthermore, TLR2 and TLR4 agonists rescued DXM-inhibited NETosis, and neither TLR2 nor TLR4 antagonists could further inhibit NETosis reduction induced by DXM, indicating that DXM may inhibit NETosis by regulating TLR2 and TLR4. In conclusion, the mechanisms of S. aureus- and PMA-induced NETosis are different. DXM decreases NETs formation independently of oxidant production and NF-κB phosphorylation and possibly via a TLR-dependent mechanism.

Allogeneic hematopoietic stem cell transplantation (HSCT) is a well-established therapeutic option for a range of inherited and acquired hematological disorders. However, graft-versus-host disease (GVHD) remains the leading cause of non-relapse mortality in allogeneic HSCT recipients. Ocular involvement occurs in up to 80% of chronic GVHD patients. In our cases, the diagnosis of vitamin A deficiency was suspected for GVHD patients. Serum vitamin A measurements were conducted to confirm clinical suspicions. Our study revealed significant decrease in serum levels of vitamin A in chronic liver GVHD patients. Although there have been many studies evaluating ocular manifestations in patients with GVHD, the present study is, to our knowledge, the first to study the relationship between vitamin A and ocular manifestations of GVHD in humans. Our data suggest that vitamin A deficiency affects the severity of ocular GVHD in adults.

The first cobalt-catalyzed direct methylation of a C(sp(2) )-H bond using dicumyl peroxide (DCP) as both the methylating reagent and hydrogen acceptor has been established. The reaction proceeded without the use of any additives, and was proven to be applicable to various amides bearing a 2-pyridinylisopropyl (PIP) directing group, providing an efficient access to o-methyl aryl amides with high functional-group tolerance. Preliminary mechanistic studies suggest a radical process would be involved in the catalytic process.

Neuroblastoma is a common solid malignant tumor of the sympathetic nervous system, which contributes to 15% of cancer-related mortality in children. The differentiation status of neuroblastoma is correlated with clinical outcome, and the induction of differentiation thus constitutes a therapeutic approach in this disease. However, the molecular mechanisms that control the differentiation of neuroblastoma remain poorly understood. The present study aimed to define whether GATA3 is involved in the differentiation of neuroblastoma cells. The results demonstrated that GATA3 is a prognostic marker for survival in patients with neuroblastoma, and that high-level GATA3 expression is associated with increased self-renewal and proliferation of neuroblastoma cells. Retinoic acid treatment led to GATA3 downregulation together with neuronal differentiation, suppression of cell proliferation and inhibition of tumorigenecity in neuroblastoma cells. These findings suggest that GATA3 is a key regulator of neuroblastoma differentiation, and provide a novel potential therapeutic strategy for the induction of neuroblastoma differentiation.